Of the numerous biological agents that may be used as weapons, the Working Group on Civilian Biodefense has identified Bacillus anthracis as one of the most serious agents. The most effective defense against these agents on a broad scale is through an aggressive vaccination: program. Currently, there is one vaccine for anthrax approved for use in the United States, produced by the BioPort Corporation in Lansing Michigan. With recent events foreshadowing broader usage of vaccines against these biological agents in the general population, there has been substantial interest in the development of DNA-based vaccines due to the potential advantages associated with this approach. Genetic immunization, rather than direct delivery of recombinant proteins is beneficial since the potential for introduction of co-purifying contaminating proteins is eliminated. Although genetic vaccines typically have utilized naked plasmid DNA (pDNA) injections into muscle tissue to generate the immune response, the issue of vaccine stability and effectiveness will probably preclude their wide spread use. We are developing a novel technology for DNA delivery. The carrier has great utility for gene delivery, particularly in the area of genetic vaccines. In this application, we propose to examine the DNA binding and expression characteristics of the conjugate in vitro and in vivo, using a series of analogs and a mammalian expression plasmid that encodes potent anthrax antigens. The goal of this project will be the generation of information on how well carrier/pDNA complexes elicit an immune response in an animal model. These results will allow the evaluation of the effect of structural changes on the binding affinity for the pDNA and the resultant effect on immune response. To aid in the development of the technology for DNA vaccines, we have established collaborations with BioPort Corporation and Dr. Blake Roessler at the University of Michigan. We feel that development of this DNA delivery technology for either oral or injectable vaccines will have an immediate impact in the area of anthrax vaccine and will also have utility for a wide variety of other vaccines.